LOCUS       Exported                6595 bp ds-DNA     circular SYN 05-APR-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6595)
  AUTHORS   Edouard Evangelisti
  TITLE     Direct Submission
  JOURNAL   Exported Jun 14, 2018 from SnapGene 4.0.5
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6595
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(323..341)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(348..364)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(505..960)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        987..1091
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1092..1952
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2123..2710
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2998..3019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3034..3064
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3072..3088
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3096..3112
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3133..3151
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    3338..3462
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(3503..3808)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(4150..4809)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(4863..4893)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(4919..5042)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        5057..5640
                     /label=Hsp70 promoter
     primer_bind     5224..5266
                     /label=Hsp70_IFF
     primer_bind     complement(5636..5675)
                     /label=Hsp70-GmR_R
     CDS             5656..6189
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     5656..5675
                     /label=GmR_F
     primer_bind     complement(6171..6220)
                     /label=Hsp70_IFR
     terminator      6376..6595
                     /label=Hsp70 terminator
ORIGIN
        1 tacgccgcct tagtaacaaa catgccaaaa gaaggaaagc tttcaccagc tttcaagttt
       61 aatcaatctt tgagagtcac aattgcaaga ataaatgaat ttttacaaca cagttcgatg
      121 cactgtgttt cgtcttgatg aaaattggac ggcaaatcga gtatcttaaa caaatctcgg
      181 cataggctgt ccattttgtg gtcgcctctt tgttggtagt ttgcagtagt ttcattagta
      241 gaagatctgc atcctcaatc tgctaaacgt cttgagcagc tatgagacga tccggggaat
      301 tggggcccga gctccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt
      361 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
      421 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
      481 ttgcgcagcc tgaatggcga atggaaattg taagcgttaa tattttgtta aaattcgcgt
      541 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
      601 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
      661 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
      721 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
      781 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
      841 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
      901 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
      961 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1021 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1081 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1141 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1201 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1261 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1321 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1381 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1441 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1501 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1561 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1621 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     1681 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     1741 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     1801 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     1861 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     1921 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1981 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2041 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2101 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2161 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2221 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2281 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2341 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2401 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2461 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2521 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2581 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2641 tcgggtttcg ccacctctga cttgagcgtc gattttgtga tgctcgtcag gggggcggag
     2701 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2761 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     2821 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     2881 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     2941 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     3001 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     3061 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     3121 cgccaagctc gaaattaacc ctcactaaag ggaacaaaag ctggtaccgg gcccattata
     3181 cctcgtattc gattaaatga tacaactcat tgccacaccg caagacggcg tcaccaccat
     3241 agacagcctt tcaagcaccc ccttgtctat gagctgccac gtcagctgcc gctcctaata
     3301 tccacccgcg tccctattcc cgtcatatgc caagctatca actttgtata gaaaagttga
     3361 acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca
     3421 gactacataa tactgtaaaa cacaacatat ccagtcacta tggtcgacct gcagactggc
     3481 tgtgtataag ggagcctgac atttatattc cccagaacat caggttaatg gcgtttttga
     3541 tgtcattttc gcggtggctg agatcagcca cttcttcccc gataacggag accggcacac
     3601 tggccatatc ggtggtcatc atgcgccagc tttcatcccc gatatgcacc accgggtaaa
     3661 gttcacgggg gactttatct gacagcagac gtgcactggc cagggggatc accatccgtc
     3721 gcccgggcgt gtcaataata tcactctgta catccacaaa cagacgataa cggctctctc
     3781 ttttataggt gtaaacctta aactgcattt caccagcccc tgttctcgtc ggcaaaagag
     3841 ccgttcattt caataaaccg ggcgacctca gccatccctt cctgattttc cgctttccag
     3901 cgttcggcac gcagacgacg ggcttcattc tgcatggttg tgcttaccga accggagata
     3961 ttgacatcat atatgccttg agcaactgat agctgtcgct gtcaactgtc actgtaatac
     4021 gctgcttcat agcatacctc tttttgacat acttcgggta tacatatcag tatatattct
     4081 tataccgcaa aaatcagcgc gcaaatacgc atactgttat ctggctttta gtaagccgga
     4141 tcctctagat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     4201 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag
     4261 caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc
     4321 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
     4381 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
     4441 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     4501 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     4561 caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca tcaggcgggc
     4621 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
     4681 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
     4741 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     4801 tttctccatt ttagcttcct tagctcctga aaatctcgac ggatcctaac tcaaaatcca
     4861 cacattatac gagccggaag cataaagtgt aaagcctggg ggtgcctaat gcggccgcca
     4921 tagtgactgg atatgttgtg ttttacagta ttatgtagtc tgttttttat gcaaaatcta
     4981 atttaatata ttgatattta tatcatttta cgtttctcgt tcaactttat tatacatagt
     5041 tgataattca catatgcgtc aatgcaattg actgtgtcct cgattatttc tattattaat
     5101 attatttcca ttaaaaaaga aatccagttc gtggcaaaat aagacaaagg tttaaataaa
     5161 aatttatcgt tattaaactc gtaacttacc ttcttcatgg ctcaacccaa tctgtatatc
     5221 gacttattta atttggttaa caaatcggtt ttcgtcgcaa ataggggatg acaactaaaa
     5281 taccaatttg tgtccttgcg acacgtcaac aggtggctgc caatatctca ggttttgagc
     5341 acacaacgct gtgattggct atctgtagat cgcttcgaga actttcggac cgacatatca
     5401 gaacgcagtc atgtccaatc cagctaacga ttggtcgaaa ccgcacttca tcgcttcatt
     5461 tcaaccaatt cccatctact aaaacgagcg tcggaatagt ctcgaacgta accccacgta
     5521 gtgggtggca gcgcccctcg tatctttcaa tcgccactcg agtctcaagt ttgctcactt
     5581 tgaaattttc catctggact acacatcaaa tctaactcaa cttttatcaa ttgaaacaca
     5641 agggccccgt ttcgcatgtt acgcagcagc aacgatgtta cgcagcaggg cagtcgccct
     5701 aaaacaaagt taggtggctc aagtatgggc atcattcgca catgtaggct cggccctgac
     5761 caagtcaaat ccatgcgggc tgctcttgat cttttcggtc gtgagttcgg agacgtagcc
     5821 acctactccc aacatcagcc ggactccgat tacctcggga acttgctccg tagtaagaca
     5881 ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc ggcttacgtt
     5941 ctgcccaggt ttgagcagcc gcgtagtgag atctatatct atgatctcgc agtctccggc
     6001 gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca tgaggccaac
     6061 gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc cgcagtggct
     6121 ctctatacaa agttgggcat acgggaagaa gtgatgcact ttgatatcga cccaagtacc
     6181 gccacctaag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     6241 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6301 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6361 ccccggatcc ccggggtcga ctagaatact tagcaattgg ttagtttgaa aattgtcaaa
     6421 gcagagtttt ttcgacaata aaatttattt tctaaatatt tgatttagcg ttgacttaat
     6481 ttgttgattc ggttgtcatc ttttgtgaag atgcccgaga gtctagtaaa tctcaagcga
     6541 aacatagtga gaaggtcact atggctggtt tgcatgagtc tgggcctatt gtgct
//
